All non-sensory pertussis toxin sensitive G protein alpha subunits have been knocked out in earlier years by gene targeting. Previously we had found that Gi2 KO mice develop ulcerative colitis (UC) in 129Sv mice but not C57BL mice, and that Go KO mice develop a turning syndrome. In collaborative studies we also noted that expression of Go-alpha blocks cAMP-induced neurite outgrowth.[unreadable] [unreadable] During the previous year we completed studies on the cerebellar foliation defect in Gi2 KO mice. We discovered that Gi3 KO mice have a developmental defect in ribcage formation and, in collaboration with Bernd Nuernberg from the University of Duessledorf, Germany, found that Gi3 KO mice fail to show insulin's inhibition of livewr autophagy. A vector has been made to generate transgenic mice that express Go-alpha only in dopaminergic neurons to test the hypothesis that Go-deficient mice circle because of lack of Go in their dopaminergic neurons. HIT and RUN (also IN and OUT) vectors were designed and genetically engineered mice have been obtained with loxP sites flanking either Gi2 alpha exons 2-3-4 or Go exon exon 6. The response of macrophages to Toll-like receptor agonsts has been chracterized.